Clinical Evaluation of Extracellular ADMA Concentrations in Human Blood and Adipose Tissue

Circulating asymmetrical dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthesis, has been proposed as a biomarker for clinical outcome. Dimethylarginine dimethylaminohydrolase (DDAH) is the main enzyme responsible for ADMA metabolism and elimination. Adipose tissue ADMA concentrations and DDAH activity and their role in diabetes and obesity have not yet been investigated. In this study, we evaluated clinical microdialysis in combination with a sensitive analytical method (GC-MS/MS) to measure ADMA concentrations in extracellular fluid. Adipose tissue ADMA concentrations were assessed before and during an oral glucose tolerance test in lean healthy subjects and subjects with diabetes (n = 4 each), and in morbidly obese subjects before and after weight loss of 30 kg (n = 7). DDAH activity was determined in subcutaneous and visceral adipose tissue obtained during laparoscopic surgery (n = 5 paired samples). Mean interstitial ADMA concentrations did not differ between study populations (healthy 0.17 ± 0.03 μM; diabetic 0.21 ± 0.03 μM; morbidly obese 0.16 ± 0.01 and 0.17 ± 0.01 μM before and after weight loss, respectively). We did not observe any response of interstitial ADMA concentrations to the oral glucose challenge. Adipose tissue DDAH activity was negligible compared to liver tissue. Thus, adipose tissue ADMA plays a minor role in NO-dependent regulation of adipose tissue blood flow and metabolism.     

Cellular Scent of Influenza Virus Infection.

Volatile organic compounds (VOCs) emanating from humans have the potential to revolutionize non‐invasive diagnostics. Yet, little is known about how these compounds are generated by complex biological systems, and even less is known about how these compounds are reflective of a particular physiological state. In this proof‐of‐concept study, we examined VOCs produced directly at the cellular level from B lymphoblastoid cells upon infection with three live influenza virus subtypes: H9N2 (avian), H6N2 (avian), and H1N1 (human). Using a single cell line helped to alleviate some of the complexity and variability when studying VOC production by an entire organism, and it allowed us to discern marked differences in VOC production upon infection of the cells. The patterns of VOCs produced in response to infection were unique for each virus subtype, while several other non‐specific VOCs were produced after infections with all three strains. Also, there was a specific time course of VOC release post infection. Among emitted VOCs, production of esters and other oxygenated compounds was particularly notable, and these may be attributed to increased oxidative stress resulting from infection. Elucidating VOC signatures that result from the host cells response to infection may yield an avenue for non‐invasive diagnostics and therapy of influenza and other viral infections.     

Rapid Total Synthesis of DARPin pE59 and Barnase

We report the convergent total synthesis of two proteins: DARPin pE59 and Bacillus amyloliquefaciens RNase (Barnase). Leveraging our recently developed fast‐flow peptide‐synthesis platform, we rapidly explored numerous conditions for the assembly of long polypeptides, and were able to mitigate common side reactions, including deletion and aspartimide products. We report general strategies for improving the synthetic quality of difficult peptide sequences with our system. High‐quality protein fragments produced under optimal synthetic conditions were subjected to convergent native chemical ligation, which afforded native full‐length proteins after a final desulfurization step. Both DARPin and Barnase were folded and found to be as active as their recombinant analogues.     

Small‐Molecule Proteomimetic Inhibitors of the HIF‐1α–p300 Protein–Protein Interaction

The therapeutically relevant hypoxia inducible factor HIF‐1α–p300 protein–protein interaction can be orthosterically inhibited with α‐helix mimetics based on an oligoamide scaffold that recapitulates essential features of the C‐terminal helix of the HIF‐1α C‐TAD (C‐terminal transactivation domain). Preliminary SAR studies demonstrated the important role of side‐chain size and hydrophobicity/hydrophilicity in determining potency. These small molecules represent the first biophysically characterised HIF‐1α–p300 PPI inhibitors and the first examples of small‐molecule aromatic oligoamide helix mimetics to be shown to have a selective binding profile. Although the compounds were less potent than HIF‐1α, the result is still remarkable in that the mimetic reproduces only three residues from the 42‐residue HIF‐1α C‐TAD from which it is derived.     

Adsorption of naphthalene and indole on F300 MOF in liquid phase by the complementary spectroscopic,kinetic and DFT studies

Metal–organic frameworks (MOFs) are the promising functional materials for adsorption in liquid phase. It is important to understand the details of chemical bonding between the aromatic and heteroaromatic adsorbates and major structural units in the MOF sorbent—metal coordinatively unsaturated sites (CUS) and organic linkers. In this paper, we report the mechanistic studies of adsorption of naphthalene and indole on F300 Basolite MOF by two complementary spectroscopic methods and density functional theory (DFT) calculations. Fluorescence spectra, the near-UV/visible diffuse reflectance spectroscopy (near-UV/VIS DRS) and DFT calculations suggest that naphthalene forms an adsorption complex with F300 MOF where naphthalene is quantum confined within the cavity of the F300, weakly electronically bound to the Fe(III) CUS, and strongly dispersively stabilized by side interactions with the benzene rings of the linker of F300. On the other hand, indole forms an adsorption complex with F300 MOF in which indole is electronically bound to the Fe(III) CUS and dispersively stabilized by the side interactions with benzene rings of the linker. Coordination bonds between indole and F300 MOF in the adsorption complex are detected by geometry optimization using the DFT method, and electronic spectra are calculated by the time-dependent-DFT method. The direct spectroscopic proof of the formation of adsorption complex with coordination bonds between indole and F300 MOF is provided by the complementary near-UV/VIS DRS spectroscopy (new absorption bands at 460–660 nm), the wavelength-dependent fluorescence spectroscopy (new fluorescence bands at 410 and 430 nm) and by the time-dependent fluorescence spectroscopy.     

Structural and Biochemical Studies of Actin in Complex with Synthetic Macrolide Tail Analogues

The actin filament‐binding and filament‐severing activities of the aplyronine, kabiramide, and reidispongiolide families of marine macrolides are located within the hydrophobic tail region of the molecule. Two synthetic tail analogues of aplyronine C (SF‐01 and GC‐04) are shown to bind to G‐actin with dissociation constants of (285±33) and (132±13) nM , respectively. The crystal structures of actin complexes with GC‐04, SF‐01, and kabiramide C reveal a conserved mode of tail binding within the cleft that forms between subdomains (SD) 1 and 3. Our studies support the view that filament severing is brought about by specific binding of the tail region to the SD1/SD3 cleft on the upper protomer, which displaces loop‐D from the lower protomer on the same half‐filament. With previous studies showing that the GC‐04 analogue can sever actin filaments, it is argued that the shorter complex lifetime of tail analogues with F‐actin would make them more effective at severing filaments compared with plasma gelsolin. Structure‐based analyses are used to suggest more reactive or targetable forms of GC‐04 and SF‐01, which may serve to boost the capacity of the serum actin scavenging system, to generate antibody conjugates against tumor cell antigens, and to decrease sputum viscosity in children with cystic fibrosis.     

MALDI-TOF MS and Morphology Studies of Thiacalixarene-Silsesquioxane Products of Oligo- and Polycondensation

For the first time condensed silsesquioxane derivatives of tetratrialkoxysilyl compounds have been characterized by MALDI-TOF mass spectrometry. Tetrasubstituted p-tert-butyl thiacalix[4]arene derivatives containing organosilicon fragments with variable stereochemistry were chosen as organosilicon compounds for polycondensation. Information obtained from mass spectra was used to deduce both the structures of oligomeric derivatives, as well as the structure of silsesquioxane framework. The morphology of formed polysilsesquioxanes was investigated by scanning and transmission electron microscopy.     

Ecological resilience and resilient cities

The urban realm is changing rapidly and becoming increasingly interconnected across continents, and across contrasting types of land covers, while at the same time facing new environmental threats and experiencing new demographic and social pressures. The urban component of the global ecosystem can be made more sustainable by incorporating the ecological understanding of resilience into the discourse. Sustainability is seen as a social, normative goal, which can be promoted using the mechanisms of ecological resilience. Ecological resilience differs from engineering resilience. Ecological resilience emphasizes the capacity of a site to adjust to external shocks and changes in controlling interactions, while engineering resilience emphasizes its ability to return to a state that existed before perturbation. Ecological resilience is particularly appropriate to urban systems, given the extent and open-ended nature of the changes and challenges they face. Adaptive processes are explored as contributions to the achievement of a successful adaptive cycle in urban socio-ecological systems. Key tools for incorporating the ecological thinking about resilience into the social discourse include landscape or patch ecology, the novel idea of the metacity, an assessment of ecological and design models, and the use of designs as experiments.     

A Comparison of the Microwave Photoconductivity Decay and Open-Circuit Voltage Decay Lifetime Measurement Techniques for Lifetime-Enhanced 4H-SiC Epilayers

This work compares the optical microwave photoconductivity decay (μPCD) and electrical open-circuit voltage decay (OCVD) techniques for measuring the ambipolar carrier lifetime in 4H-silicon carbide (4H-SiC) epitaxial layers. Lifetime measurements were carried out by fabricating P⁺/intrinsic/N⁺ (PiN) diodes on 100-μm-thick, 1 × 10¹⁴ cm^?3 to 4.5 × 10¹⁴ cm^?3 doped N-type 4 H-SiC epilayers, and measuring the lifetime optically using μPCD prior to metallization, then electrically using OCVD after contact deposition. Both as-grown epilayers as well as epilayers with improved lifetime (via thermal oxidation) were measured using both techniques. The observed ambipolar lifetime was improved from 1.4 μs on an unenhanced wafer to 4 μs on a wafer enhanced through the oxidation process as measured by μPCD. Little difference was observed between the μPCD and OCVD measurements on the unenhanced wafer; the ambipolar lifetime on the enhanced wafer measured by OCVD was approximately 5.5 μs, or 1.5 μs higher than the μPCD measurement. Continuous evaluation of the OCVD transient waveform was necessary due to the high lifetime in the enhanced wafer; shunt resistances included to discharge the P⁺/N junction capacitance were found to damp the OCVD response and yield low values for the measured lifetime. Simulation of the μPCD measurement including various surface recombination conditions yielded a good match to experimentally observed μPCD measurements for high values of the surface recombination velocity. The OCVD lifetime measurement technique is expected to yield measured lifetime values closer to the physical value due to its independence from surface conditions, provided that the experimental conditions are appropriately chosen.